Centrifugal precitation chromatography has been successfully applied to purify an enzyme called carotenoid cleavage enzymes from Japanese tea leaves (Camellia sinensis) with high purity. Preliminary experiments were performed to determine the critical solvent composition and concetration of the solvent composed of acetone and ethanol buffered with Tris so that the native enzymatic activity is preserved. Various other separation parameters such as flow rate, rotation speed, and sample load were also optimized by a series of pilot experiments. Subsequently, this newly developed separation protocol was used for the separation of active catotenoid cleavage enzymes from tea leaves, and the obtained fractions were screened for enzymatic acitivity. The measured enzymatic activity was found to be much higher than that obtained from the classical protein purification protocols. The purity of the enzyme was excellent and the samples could be directly used for structural elucidation. In conclusion centrifugal precipitation chromatography is proved to be a powerful technique for the isolation of active enzymes from natural sources. These results have been presented at the 5th International Conference on Countercurrent Chromatography which was held in Rio de Janeiro, Brazil on July 26 - 29, 2008.